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PMID: 9414322 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The complete set of predicted genes from Saccharomyces cerevisiae in a readily usable form.

Genome research ·Vol. 7 ·No. 12 ·1997-12-00 ·Pages 1169-73

Hudson JR, Dawson EP, Rushing KL, Jackson CH, Lockshon D, Conover D, Lanciault C, Harris JR, Simmons SJ, Rothstein R, Fields S

Abstract

Nearly all of the open reading frames (ORFs) of the yeast Saccharomyces cerevisiae have been synthesized by PCR using a set of approximately 6000 primer pairs. Each of the forward primers has a common 22-base sequence at its 5' end, and each of the back primers has a common 20-base sequence at its 5' end. These common termini allow reamplification of the entire set of original PCR products using a single pair of longer primers-in our case, 70 bases. The resulting 70-base elements that flank each ORF can be used for rapid and efficient cloning into a linearized yeast vector that contains these same elements at its termini. This cloning by genetic recombination obviates the need for ligations or bacterial manipulations and should permit convenient global approaches to gene function that require the assay of each putative yeast gene.

MeSH Terms
DNA Primers DNA, Fungal/analysis Genes, Fungal Genome, Fungal Open Reading Frames/genetics Polymerase Chain Reaction Saccharomyces cerevisiae/genetics Transformation, Genetic
Chemicals
DNA Primers DNA, Fungal
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Hudson J R
Research Genetics Inc., Huntsville, Alabama 35801, USA.
Dawson E P
Rushing K L
Jackson C H
Lockshon D
Conover D
Lanciault C
Harris J R
Simmons S J
Rothstein R
Fields S
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Article Info
Journal
Genome research
Abbr.
Genome Res
ISSN
1088-9051
Published
1997-12-00
Pages
1169-73
Language
English
Region
United States
NLM ID
9518021
PMCID
PMC310675
Subset
IM
Grants
NIGMS NIH HHS · GM54415 · United States
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